COPB2 gene silencing inhibits colorectal cancer cell proliferation and induces apoptosis via the JNK/c-Jun signaling pathway

PLoS One. 2020 Nov 19;15(11):e0240106. doi: 10.1371/journal.pone.0240106. eCollection 2020.

Abstract

Objectives: Colorectal cancer (CRC) is one of the most common malignant human tumors. It is associated with high morbidity and mortality rates. In recent years, tumor gene therapy has emerged as a promising new approach for colorectal cancer therapy. Herein, we identify and analyze the role of COPB2 (coatomer protein complex, subunit beta 2) in proliferation and apoptosis of CRC cells.

Methods: To investigate the role of COPB2 in the proliferation and apoptosis of CRC cells, a shCOPB2 vector and a shCtrl vector were constructed for transfection into RKO and HCT116 cells. Cells proliferation was subsequently measured via cell counting kit-8 (CCK8) assay and Celigo cell counting assay. Apoptosis was measured via flow cytometry. The activity level of Caspase 3/7 was measured. Finally, the level of several JNK/c-Jun apoptosis pathway-related proteins were measured to characterize the mechanism of apoptosis.

Results: Our results showed that the proliferation rate was decreased and the apoptosis rate was increased in shCOPB2-treated RKO and HCT116 cells compared to those in controls. After the silencing of COPB2, JNK/c-Jun signal pathway activation was increased, the expression levels of apoptosis pathway-related proteins, such as Bad, p53 and Caspase 3, were also increased.

Conclusion: COPB2 gene silencing can inhibit RKO and HCT116 cells proliferation and induce apoptosis via the JNK/c-Jun signaling pathway.

MeSH terms

  • Cell Line, Tumor
  • Cell Movement / drug effects
  • Cell Proliferation / drug effects
  • Cell Survival / drug effects
  • Coatomer Protein / antagonists & inhibitors
  • Coatomer Protein / genetics*
  • Colorectal Neoplasms / drug therapy
  • Colorectal Neoplasms / genetics*
  • Colorectal Neoplasms / metabolism
  • Gene Expression Regulation, Neoplastic / drug effects
  • Gene Silencing
  • HCT116 Cells
  • HT29 Cells
  • Humans
  • MAP Kinase Signaling System
  • Proto-Oncogene Proteins c-jun / metabolism
  • RNA, Small Interfering / pharmacology*
  • Survival Analysis
  • Up-Regulation / drug effects*

Substances

  • COPB2 protein, human
  • Coatomer Protein
  • JUN protein, human
  • Proto-Oncogene Proteins c-jun
  • RNA, Small Interfering

Grants and funding

The authors received no specific funding for this work.